Hyaluronic acid and chondroitin sulfate based hydrolyzed collagen type II and method of making same

ABSTRACT

Hydrolyzed collagen type II powder compositions for inducing cartilage formation in an individual, method of preparing the compositions and use of the compositions in treating connective tissue disorder, replenishing skin viscoelasticity. The compositions are administered through an orally ingestible delivery medium for absorption into the gastrointestinal tract. The compositions are administered through a topical delivery medium for absorption into a dermis of the individual.

CROSS-REFERENCE TO RELATED APPLICATIONS

(The present application is a continuation application of U.S.application Ser. No. 10/008,012 entitled HYALURONIC ACID AND CHONDROITINSULFATE BASED HYDROLYZED COLLAGEN TYPE II AND METHOD OF MAKING SAMEfiled Nov. 13, 2001 now U.S. Pat. No. 6,780,841)

STATEMENT RE: FEDERALLY SPONSORED RESEARCH/DEVELOPMENT

(Not Applicable)

BACKGROUND OF THE INVENTION

The present invention relates to hydrolyzed collagen type II powdercompositions for inducing cartilage formation in an individual, a methodof preparing the compositions and the use of the compositions intreating connective tissue disorder and replenishing skinviscoelasticity.

Studies have shown that collagen is a complex structural protein whichprovides strength and flexibility to skin, hair and nails. Collagen isan essential and major component of muscles, tendons, cartilage,ligaments, joints and blood vessels in the human body. There are threemain types of collagen: I, II and III. Types I and III are primarilyfound in skin, tendon and bone. In contrast, type II is foundpredominantly in articular cartilage. Collagen is an unusual protein, inthat the proportion of glycine residues is nearly one-third, which isunusually high in comparison to other typical proteins. Proline is alsopresent to a much greater extent in collagen than in most otherproteins. Moreover, collagen contains two amino acids, 4-hydroxyprolineand 5-hydroxylysine, that are found in very few other proteins. Theamino acid sequence of collagen is remarkably regular, nearly everythird amino acid being glycine. In addition, the sequenceglycine-proline-hydroxyproline recurs frequently. In contrast, globularproteins rarely exhibit regularities in their amino acid sequences(Stryer, L., Biochemistry, Third Edition, W.H. Freeman and Co., NewYork, 1988, pp. 262).

In 1986, collagen was sold for the first time in the United States foruse as a food supplement. Collagen (a mixture of Types I and III) wasextracted from calf skin tissue, hydrolyzed and prepared in powderedform for use as a dietary supplement. The composition was sold under thename “Hydrolyzed Collagen Beauty Supplement™” (Smarter Nails & Hair,Inc., Newport Beach, Calif.). In 1987, “Hydrolyzed Collagen BeautySupplement Tablet™” (Smarter Nails & Hair, Inc., Newport Beach, Calif.)was sold which comprised collagen powder and 10 mg vitamin C compressedinto 1,000 mg tablets.

U.S. Pat. No. 4,804,745 to Koepff et al. discloses agents containingcollagen peptides produced by enzymatic hydrolysis for the treatment ofdegenerative joint diseases. These peptides can be obtained from animalskin, animal bones and other sufficiently purified connective tissue andhave average molecular weights of between 30 and 45 kilodaltons.

U.S. Pat. No. 5,399,347 to Trentham et al. and Trentham et al. (Science261:1727–1729, 1993) disclose the effective treatment of rheumatoidarthritis (RA) with water-soluble whole chick collagen type II orbiologically active peptides derived therefrom. The mechanism by whichthe effect is believed to occur is via oral tolerization toautoantigens.

U.S. Pat. No. 5,364,845 to Henderson discloses a therapeutic compositionand method for the protection, treatment and repair of connective tissuein mammals. This composition comprises glucosamine, chondroitin sulfateand manganese ascorbate. U.S. Pat. No. 5,587,363 to Henderson disclosesa therapeutic composition and method for the protection, treatment andrepair of connective tissue in mammals which includes aminosugars andglycosaminoglycans.

Another well known substance which aids in the rejuvenation andtreatment of such ailments as connective tissue disorder is a substanceknown as Hyaluronic Acid (also known as “Hyaluronan” or “HA”). It iswell known that the human body naturally contains such HA, as it isfound in several parts of the body such as the soft connective tissue,the vitreous body of the eye, hyaline cartilage, synovial joint fluid,the dermis, and the epidermis. Within these parts of the body, HA actsas a lubricant between connective tissues of the skin, protects thejoints by providing shock-absorption, and helps the body retain skinmoisture. Over time however, as the body ages, the amount of HA presentin the body deteriorates and the body may eventually develop one ofseveral health problems, in part due to a decreased presence of HA. Thiseffect is particularly apparent for those who are over the age of 50.Generally, the skin loses viscoelasticity, and wrinkles form ultimatelyas a result of this deficiency.

Due to the fact that HA is found in several different parts of the body,such a decrease in HA levels is associated with a great variety ofdisorders and ailments. For example, osteoarthritis patients experiencedecreased levels of HA in their synovial fluid. This has a detrimentaleffect on the joints because HA is primarily responsible for thelubricating and shock-absorbing effects of the synovial fluid. For thisreason, researchers have theorized that the replacement of such lost HAmay help osteoarthritis sufferers to rebuild damaged cartilage and toregain joint movement.

However, HA is not found exclusively in the human body. As a matter offact, HA is found in most animals. For this reason, there has beensubstantial research conducted to extract HA from other sources so thathumans may replenish the levels of HA that are lost over time. HA may beextracted from several sources such as from micro-organisms(Streptococcal cultures) through a fermentation process or from theisolation HA from rooster combs. Of these major sources, rooster combsproduce HA having higher molecular weights. HA having a high molecularweight is too large to penetrate into the skin and the bloodstream andis therefore limited in its application. Generally, high molecularweight HA is used for eye surgery and cosmetics, specifically forcreating a viscoelastic film to retain skin moisture and to blockforeign substances.

For Osteoarthritis (“OA”) patients, one method of relieving joint painis via a therapy known as viscosupplementation. Viscosupplementationrequires an intra-articular injection of HA to alleviate the painassociated with OA and allows the patient to regain function of theaffected joints. The end result of such treatment is that the HA has ananti-inflammatory effect. Alternatively, the patient may useintra-articular glucocorticoid injections. The benefit of bothinjections are somewhat comparable but the common disadvantage to boththerapies is the invasive nature of the injections.

Another constituent which alleviates pain in the joints and helpsrebuild connective tissue is an endogenous Glycosaminoglycan (“GAG”)called Chondroitin Sulfate (“CSA”) which is composed of GlucuronicAcid+N-acetyl-D-galactosamine+Glucosamine Sulfate. Generally, CSA is abiological polymer derived from connective tissue. There are three typesof Chondroitin Sulfate: Chondroitin Sulfate A (“CSA-A”) (shown in FIG.2) which is also known as chondroitin 6-sulphate, Chondroitin Sulfate B(“CSA-B”) (shown in FIG. 3) which is also known as dermatan sulphate,and Chondroitin Sulfate C (“CSA-C”) (shown in FIG. 4) which is alsoknown as chondroitin 4-sulphate. The ratio of CSA-A to CSA-C declines intandem with the progression of the aging process as does the content ofCSA-B in the Dermis of the skin. In this respect, CSA levels aregradually depleted and such a decrease in CSA levels has a similareffect to a decrease in HA levels in that the shock-absorption andlubrication qualities are reduced. Thus, like HA, CSA is generally animportant GAG for the maintenance of connective tissue.

Specifically, CSA-A has proven to be an effective anti-inflammatory thatimproves blood circulation, prevents Ischemic Heart Disease, reducesincidences of heart attacks, reduces incidences of strokes, and is alsoeffective in supporting connective joint tissue. Insofar as ChondroitinSulfate helps prevent heart disease, this phenomenon is due to theinherent antithrombogenic or anticoagulant properties which preventabnormal blood clots and reduce the incidence of strokes. CSA-B is foundin the dermis of the skin and is also one of the constituentsresponsible for maintaining viscoelasticity of the skin. CSA-C inhibitsElastase, which is an enzyme that progressively degrades cartilageduring the onset of Osteoarthritis.

Currently, Chondroitin Sulfate may be naturally ingested via seafood.For example, mussels, oysters and shark cartilage naturally containChondroitin Sulfate such that ingesting these types of foods may helpreplenish lost Chondroitin Sulfate over time. However, it is impracticalto continually ingest seafood on a regular basis because some people areallergic to particular types of seafood, seafood is relativelyexpensive, and some simply do not like the taste of seafood.Additionally, several Chondroitin Sulfate nutritional supplements arebeing sold currently for treating connective joint tissue. Most of thesesupplements derive the Chondroitin Sulfate from bovine cartilage orvelvet deer antler. In these forms, Chondroitin Sulfate is generally adifficult compound to digest in the gastrointestinal tract and the typeof Chondroitin Sulfate derived from such animals is generally not veryeffective in absorbing into the bloodstream.

There is a need for non-invasive therapies in treating OA and otherconnective tissue disorders which may be treated with HA and a need forcompositions capable of promoting repair of damaged connective tissue.The present invention addresses this need.

BRIEF SUMMARY OF THE INVENTION

One embodiment of the present invention is hydrolyzed collagen type II,the hydrolyzed collagen having an average molecular weight of betweenabout 50 and 10,000 daltons. Preferably, the hydrolyzed collagen type IIhas an average molecular weight of about 5,500 daltons. In one aspect ofthis preferred embodiment, the collagen is obtained from chicken sternalcartilage. The hydrolyzed collagen type II preferably includes at least20% depolymerized chondroitin sulphate and at least 10% hyaluronic acid.The collagen may be formed into an orally ingestible delivery medium ora topical delivery medium. The collagen is a low molecular weight,thereby allowing the orally ingestible delivery medium to readily absorbinto the gastrointestinal tract of an individual while the topicaldelivery medium readily absorbs into the dermis of an individual.

The present invention also provides a method of inducing cartilageformation in an individual with a connective tissue disorder, comprisingorally administering to the individual an effective dailycartilage-inducing amount of hydrolyzed collagen type II. The connectivetissue disorder includes degenerative joint diseases, joint defects,osteoarthritis, polychondritis, vascular disease and cartilage injuries.Preferably, the effective daily amount is between about 500 and 5,000mg. More preferably, the effective daily amount is between about 1,000and 4,000 mg. Most preferably, the effective daily amount is betweenabout 2,000 and 3,000 mg.

Another embodiment of the invention is a method of replenishinghyaluronic acid and chondroitin sulfate, restoring skin viscoelasticity,retaining skin moisture, healing wounds and improving the overallappearance of skin comprising topically administering to an individual,or other animal, a selectable amount of chicken sternalcartilage-derived material comprising hydrolyzed collagen type II havingan average molecular weight of between about 50 and about 10,000daltons. In this embodiment, the patient is given the option of applyingas much or as little as desired to portions of the body as theindividual sees fit (i.e., the eyes).

Still another embodiment of the invention is a method of preparinghydrolyzed collagen type II powder, comprising the following steps:cutting fresh chicken sternal cartilage to within not less than about 2mm of the bone; suspending the cartilage in an aqueous solution;treating said cartilage with a proteolytic enzyme to form a hydrolysate,said proteolytic enzyme being capable of hydrolyzing collagen type II tofragments having an average molecular weight of between 50 to 10,000daltons; sterilizing the hydrolysate; filtering the hydrolysate;concentrating the hydrolysate; and drying the hydrolysate to form acollagen type II powder. The method may further comprise the step offreezing the cartilage after the cutting step. Preferably, the aqueoussolution is water. Advantageously, the enzyme is papain, ficin orbromelain. In one aspect of this preferred embodiment, the sterilizingstep comprises heating the hydrolysate at 95° C. for about 30 minutes.Preferably, the drying step comprises spray drying. Preferably, the pHof the suspending and treating steps is between about 4 and 8.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic diagram of the process for preparing thehydrolyzed collagen type II powder of the invention.

FIG. 2 is a molecular diagram showing Chondroitin Sulphate-A.

FIG. 3 is a molecular diagram showing Chondroitin Sulphate-B.

FIG. 4 is a molecular diagram showing Chondroitin Sulphate-C.

FIG. 5 is a molecular diagram showing Hyaluronic Acid.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a hydrolyzed, denatured collagen type IIcomposition, method for preparing the composition and use of thecomposition in the prevention, treatment and repair of cartilagedefects. The method involves cutting fresh sternal cartilage fromchicken carcasses and removing all meat therefrom. The sternal cartilageis cut, leaving a space of about two millimeters from the bone so as tonot remove any bone fragments. This is critical to the purity of thefinal product because it avoids contamination of collagen type II withtypes I and III found in bone. Collagen type II contains the greatestconcentration of proteoglycans, which help rejuvenate connective tissuejoints. Therefore, maintaining a pure collagen type II compound ensuresthat the highest concentration of such proteoglycans. The fresh sternalcartilage is then promptly frozen and the remains of the chicken carcassare discarded. It is exclusively the sternal cartilage that is used forpreparing the collagen type II powder. The chicken sternal cartilage isprocessed according to good manufacturing procedures (GMP).

Other contemplated sources of collagen type II are mammalian cartilage(e.g. bovine, porcine and avian) and shark fins. However, such forms ofcollagen type II are generally less effective in absorbing into thegastrointestinal tract. On the other hand, the present inventionnaturally contains depolymerized chondroitin sulfate. This particularform of chondroitin sulfate derived from the chicken sternal cartilageabsorbs into the gastrointestinal tract approximately three to fivetimes better than traditional bovine derived forms which may not bedepolymerized. In this respect, the depolymerized chondroitin sulfate isbroken down into its individual monomers for an increased ability toabsorb into the gastrointestinal tract, thereby increasing the bloodconcentrations of glycosaminoglycans rich in hydrolyzed collagen typeII. Preferably, the present invention includes 20–25% depolymerizedchondroitin sulfate, a high concentration of this easily absorbableconstituent. However, this depolymerized chondroitin sulfate alsoprovides for absorption into the body via other means. For example, thedepolymerized chondroitin sulfate in the present invention may absorbinto the skin when applied topically to replenish CSA-C in the dermis.

Similarly, the hyaluronic acid present in the collagen replenishes lostlevels of such hyaluronic acid in the body by supplementing the synovialjoint fluid, cartilage, eyes, skin tissue, connective tissue, nails, andother parts of the body which gradually lose hyaluronic acid over time.The collagen having the hyaluronic acid also provides structural supportto joints, promotes new cartilage synthesis, helps heal wounds and isuseful in a myriad of other applications where hyaluronic acid andchondroitin sulfate is generally found within the human body, or animalbody. Advantageously, because the collagen is a low molecular weight,the hyaluronic acid contained in the collagen is capable of absorbinginto the gastrointestinal tract via oral delivery mediums and can absorbinto the skin when applied topically. Thus, hyaluronic acid is alsocapable of reaching the corium layers of the dermis via oral ingestion.In this respect, the hydrolyzed collagen type II and its unique lowmolecular weight properties allow for the hyaluronic acid, chondroitinsulfate, glucosamine sulfate, cartilage matrix glycoprotein (CMGP), andother glucosaminoglycans to all absorb into the gastrointestinal tractvia oral delivery mediums and into the skin via topical deliverymediums.

The production of hydrolyzed collagen type II in powdered form is shownin FIG. 1. Whole cartilage is suspended in an aqueous solution,preferably water for about one hour at about 35° C. at a pH of betweenabout 4 and 8. In a preferred embodiment, the pH is between about 6 and7. In a more preferred embodiment, the pH is about 6.5. The water isremoved, and the cartilage is incubated with one or more proteasesobtainable from a natural source (i.e. papain, ficin, bromelain) forbetween about 2 and 10 hours, preferably about 6 hours, at about 35°C.–55° C. at a pH of between about 4 and 8 to form a hydrolysate. The pHwill depend on the pH optimum of the particular enzyme(s) used for thehydrolysis and are well known to one of ordinary skill in the art. Thehydrolysate is then sterilized for about 30 minutes at a temperaturebetween about 95° C. and 105° C. The sterilized hydrolysate is filteredthrough diatomaceous earth, concentrated, preferably under vacuum, driedto form a powder and packed. Other filtration methods are contemplated,including vacuum filtration. In a preferred embodiment, the hydrolyzedcollagen type II is spray dried using a size 56 pressure nozzle into aheat tunnel. The final particle size and mesh are adjusted to 0.46 g/cc,yielding a fine powder. The powder is packed in a 40 kg drum with aplastic bag liner. The powder is water soluble.

The average molecular weight of the final product is between 50 and10,000 daltons, preferably 5,500 daltons. The moisture content isbetween 5% and 7%. The final product is high in mucopolysaccharides,particularly chondroitin sulfate and glucosamine sulfate. Suchmucopolysaccharaides are major constituents in providingshock-absorption and lubrication in the connective tissue joints. Theproduct has 375 calories per 100 grams, contains 67% protein (12.1%total nitrogen), 18% carbohydrate and 0.1% fat. The amino acidcomposition of the hydrolysate differs substantially from typicalcollagens and is shown in Table 1. Hydroxyproline is low, hydroxylysineis absent and tryptophan is low. The low molecular weight and high aminoacid composition promote optimal assimilation of the peptides.

TABLE 1 Amino acid composition of hydrolyzed collagen type II Amino acidg/100 g product arginine 4.42 histidine 2.05 isoleucine 1.90 leucine4.20 lysine 3.54 methionine 1.38 phenylalanine 2.14 threonine 2.60tryptophan 0.37 alanine 4.51 asparagine/aspartic acid 5.29 cystine 0.46glutamine/glutamic acid 8.75 glycine 8.93 hydroxyproline 3.90 proline5.25 serine 2.45 tyrosine 1.16 valine 2.43

A test analysis of the hydrolyzed collagen yields results which indicatethat at least 20% of depolymerized chondroitin sulfate is present in thecollagen. Additionally, at least 10% of hyaluronic acid is also foundtherein. However, because the collagen is extracted from a naturalsource (i.e. chicken sternum), the actual percentage of depolymerizedchondroitin sulfate and hyaluronic acid may vary. The highestconcentration of depolymerized chondroitin sulfate and hyaluronic acidobtainable is preferred.

TABLE 2 Test Results of Hydrolyzed Collagen Type II SpecificationsTypical analysis Purity    100% Solubility Water-Soluble ChondroitinSulfate ~20.0% Hyaluronic Acid ~10.0% Other Proteoglycans  ~3.0%Collagen Type II Protein ~62.7% Nitrogen   12.1% Fat    0.0% Loss onDrying   <8% Heavy Metals <5 ppm Ash    8.3% Calcium  530 mg/100 gmsMagnesium 161 mg/100 gms Potassium 1961 mg/100 gms MicrobiologicalAnalysis Standard Plate Count <1000 gms E. Coli Negative SalmonellaNegative Physical Analysis Appearance Off white/yellow fine powder OdorCharacteristic Particle Size >100 mesh

When taken orally by an individual with a connective tissue disorder,hydrolyzed collagen type II helps build cartilage and significantlyimproves the disorder. The lost levels of HA, CSA-A, CSA-B, and CSA-C inthe joints are replenished via ingestion into the gastrointestinaltract. Furthermore, oral ingestion of the hydrolyzed collagen type IIhelps contribute to the wound healing process due to thechondroprotective nature of the collagen's composition.

“Oral” administration includes oral, enteral or intragastricadministration. The hydrolyzed collagen type II of the invention can beused to treat, for example, degenerative joint diseases (i.e. rheumatoidarthritis), joint defects, osteoarthritis, polychondritis, vasculardisease, cartilage injuries, silicone poisoning due to ruptured breastimplants, autoimmune diseases involving connective tissue autoantibodies(i.e. rheumatoid arthritis), progressive myopia, Menier's disease andany other connective tissue disorder which would benefit from increasedsynthesis of cartilage. The hydrolyzed collagen type II alsosignificantly reduces sun-induced skin wrinkles.

By extracting HA from chicken sternal cartilage, a low molecular weightHA may be obtained by hydrolysis after such extraction. This distinctionis crucial because the beneficial therapeutic activity of HA is mostlydependent upon the molecular weight of HA. Due to the low molecularweight of the HA found in the hydrolyzed collagen type II, thehydrolyzed collagen type II readily absorbs into the gastrointestinaltract and allows the rejuvenating constituents of HA and CSA to restoreviscoelasticity to the skin, protect connective tissues, promotecartilage synthesis, retain skin moisture, heal wounds and improve theoverall appearance of skin. In its oral form, the hydrolyzed collagentype II is capable of reaching the corium layers of the skin whichtypically cannot be reached by topical applications.

For oral administration as a nutritional supplement, therapeutic orprophylactic agent, the hydrolyzed collagen type II of the invention maybe provided as a dispersible powder or granule, tablet, aqueous or oilsuspension, emulsion, hard or soft capsule, syrup or elixir.Advantageously, the ability to provide the hydrolyzed collagen type IIin such a variety of forms allows the invention to be used by itself orin conjunction with several different applications.

The low molecular weight of the hydrolyzed collagen type II allows thepowder form to be integrated into topical gels. In this respect, the gelmay be applied directly to the skin and ultimately, the collagen readilyabsorbs into the skin. Absorption directly into the skin provides theadvantage of moisturizing the skin, maintaining fluid balance in thecells of the skin and providing a thin layer of viscoelastic protectivefilm which keeps foreign substances from attacking the skin.Additionally, the film retains water and moisturizes the skin in muchthe same way that hyaluronic acid does in the intercellular matrix ofdermal connective tissue. Ultimately such application helps the skinappear younger, reduces the effects of aging upon the skin, andaccelerates the wound healing process when the collagen is applied towounds. Additionally, topical application provides the advantage of atargeted solution to where the consumer needs it the most. For example,the topical collagen user's predominate purpose may be to rejuvenate theskin and restore viscoelasticity to improve the overall appearance ofthe skin. Such a user may then apply the topical collagen to the desiredparts of the body, such as under the eyes, to concentrate onrejuvenating that particular area. A topical form may also be used foradministering the collagen to wounds for reducing the appearance ofscarring and accelerating the overall wound healing process. On theother hand, an orally ingestible form provides the advantage of allowinga total body rejuvenation wherein the collagen restores hyaluronic acidand chondroitin sulfate constituents to parts of the body which aredeficient in these chemicals.

As a cosmetic, the collagen may be integrated with existing facialcosmetics, such as foundation, concealer, and lipstick as an additivewhich works invisibly to replenish the lost constituents. In thisrespect, the cosmetic user may apply the daily cosmetics as usualwithout the requirement of an additional moisturizer due to thecollagen's ability to help retain natural skin moisture. Furthermore, itis contemplated that the collagen may be used in conjunction with othertypical consumer products where skin moisturizing is also useful. Forexample, some soaps include a moisturizer as an ingredient to counteractany drying effects detergents may have on the skin. The collagen may beused with a moisturizer to also counteract such effects whilesimultaneously replenishing lost levels of HA and CSA.

The hydrolyzed collagen type II may serve as an ingredient of a productmedication manufactured to facilitate healing of wounds and injuries. Inthis regard, such product medication, which incorporates the hydrolyzedcollagen type II as one of its ingredients, may be directly medicated onareas of skin where the wound occurs. Additionally, the collagen may beintegrated into an orally ingestible form of wound healing medication byabsorbing into the gastrointestinal tract. Of course, sufficientquantity should be applied to promote a healing process of the wound.

Specifically, the hydrolyzed collagen type II of the present inventionmay be conformed to provide topical applications so as to acceleratehealing of wounds and injuries associated with the diseases/disordersdescribed above (e.g., cartilage injuries, hepatitis, decubititis). Inthis respect, when a wound heals, the fibroplasts secreteglycosaminoglycans such that a hydrophilic matrix is formed. Thehydrophilic property of the hydrolyzed collagen may then provide agreater degree of epithelialization. The increased presence of suchglycosaminoglycans assists the fibroblasts which are continually movingand replicating during the wound healing process. Thus, the overallprocess for accelerating the healing of cutaneous wounds is acceleratedby applying the collagen to the wound. In the same respect, the sameproperties of the collagen assist in plastic surgery applications forburn victims. Applying the collagen to wounds, skin graft sites, andskin donor sites may reduce inflammation, reduce the incidence ofscarring, and also increase the overall healing process.

Compositions intended for oral use may be prepared according to anymethod known in the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more of thefollowing agents: sweeteners, flavoring agents, coloring agents andpreservatives. The sweetening and flavoring agents will increase thepalatability of preparation. Tablets containing the hydrolyzed collagentype II in admixture with non-toxic pharmaceutically acceptableexcipients suitable for tablet manufacture are acceptable. Suchexcipients include inert diluents such as calcium carbonate, sodiumcarbonate, lactose, calcium phosphate or sodium phosphate; granulatingand disintegrating agents, such as corn starch or alginic acid; bindingagents such as starch, gelatin or acacia; and lubricating agents such asmagnesium stearate, stearic acid or talc. Tablets may be uncoated or maybe coated by known techniques to delay disintegration and absorption inthe gastrointestinal tract and thereby provide a sustained action over alonger period of time. For example, a time delay material such asglyceryl monostearate or glyceryl distearate alone or with a wax may beemployed. The use of enteric coatings is also contemplated.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample calcium carbonate, calcium phosphate or koalin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions may contain the hydrolyzed collage type II of theinvention in admixture with excipients suitable for the manufacture ofaqueous suspensions. Such excipients include suspending agents,dispersing or wetting agents, one or more preservatives, one or morecoloring agents, one or more flavoring agents and one or more sweeteningagents such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oil suspension maycontain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth above, and flavoringagents may be added to provide a palatable oral preparation. Thesecompositions may be preserved by an added antioxidant such as ascorbicacid. Dispersible powders and granules of the invention suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Additional excipients,for example sweetening, flavoring and coloring agents, may also bepresent.

Syrups and elixirs may be formulated with sweetening agents, such asglycerol, sorbitol or sucrose. Such formulations may also contain ademulcent, a preservative, a flavoring agent and/or a coloring agent.

The hydrolyzed collagen type II powder may be mixed with otheringestible forms and consumed in solid, semi-solid solution, suspensionor emulsion form. It may also be mixed in conjunction or alternativelywith pharmaceutically acceptable carriers, flavor enhancers, water,suspending agents and emulsifying agents. In a preferred embodiment, thehydrolyzed collagen type II powder is mixed with a citrus juice such asorange, grapefruit or tangerine due to the promotion of connectivetissue formation by ascorbic acid. The hydrolyzed collagen may also beprovided in tablet form in admixture with ascorbic acid.

For use as a nutritional supplement, prophylactic or therapeutic agent,the hydrolyzed collagen is orally administered in a daily dosage ofbetween about 500 mg and 5,000 mg. More preferably, it is administeredin a daily dosage of between about 2,000 mg and 4,000 mg. Mostpreferably, it is administered in a daily dosage of between about 2,000and 3,000 mg per day. The hydrolyzed collagen type II powder may beformulated into tablets which range from 300 mg to 1,000 mg per tablet.In a preferred embodiment, the hydrolyzed collage type II powder isformulated into 500 mg tablets and 4–6 tablets are taken daily. Inanother preferred embodiment, the tablets are taken on an empty stomachwith a beverage containing vitamin C.

In another preferred embodiment, the hydrolyzed collagen type II powderis mixed with water or a citrus juice prior to ingestion. Thepreparations described above can be taken indefinitely by individualsaffected by connective tissue disorders or by healthy individuals as apreventative agent. If desired, an individual with such a disorder cantake the preparation until no further improvement is noted in thedisorder.

It is contemplated that application of the medication may beaccomplished by wiping using sterile gloves, applying with appropriatedevices, spray, painting, or other means to produce a continuousadhesive film on the wound. If additional support for the wound isrequired, bandages, splints, other medication dispensing devices,prosthetic devices or any other devices may be applied in conjunctiontherewith.

In addition, the hydrolyzed collagen type II of the present inventionmay further be adhered as a preventive remedy. For instance, like manyother ingredients in the marketplace, the hydrolyzed collagen type IIcan be subjected to cosmetic applications used for anti-aging purposes.More particularly, as the hydrolyzed collagen type II can help reducesun-induced skin wrinkles, it may therefore be integrated into ananti-aging cosmetic as one of its essential ingredients. As such, thehydrolyzed collagen type II of the present invention can be utilized ina variety of exemplary manners due to its primary inherentcharacteristic of inducing cartilage formation.

The above detailed description of the invention is set forth solely toassist in understanding the invention. It is to be understood thatvariations of the invention, including all equivalents now known orlater developed, are to be considered as falling within the scope of theinvention, which is limited only by the following claims.

1. A composition and a corresponding test analysis, wherein the composition is derived from an animal product having (a) hydrolyzed collagen type II having an average molecular weight of between about 50 and about 10,000 daltons and (b) a measurable amount of hyaluronic acid; and the test analysis shows that the hyaluronic acid comprises at least 10% of the composition.
 2. The composition and corresponding test analysis of claim 1, wherein the composition further comprises at least 20% chondroitin sulfate.
 3. The composition and corresponding test analysis of claim 1, wherein the animal product comprises a sternum.
 4. The composition and corresponding test analysis of claim 1, wherein the animal product comprises tissue of a bird.
 5. The composition and corresponding test analysis of claim 4, wherein the bird is a chicken.
 6. The composition and corresponding test analysis of claim 1, wherein the animal product comprises a chicken sternum.
 7. The composition and corresponding test analysis of claim 1, wherein the animal product comprises chicken sternum cartilage.
 8. The composition and corresponding test analysis of claim 1, wherein the animal product comprises at least one of hyaline and articular cartilage.
 9. The composition and corresponding test analysis of claim 1, wherein the animal product comprises tissue of at least one of a cow, a pig and a shark).
 10. The composition and corresponding test analysis of claim 1, wherein at least same of the composition is heat hydrolyzed.
 11. The composition and corresponding test analysis of claim 1, wherein the collagen type II is enzymatically hydrolyzed.
 12. The composition and corresponding test analysis of claim 1, wherein the hyaluronic acid is enzymatically hydrolyzed.
 13. The composition and corresponding test analysis of claim 1, further comprising chondroitin sulfate that is enzymatically hydrolyzed.
 14. The composition and corresponding test analysis of claim 1, wherein the composition is at least partially dehydrated.
 15. The composition and corresponding test analysis of claim 1, wherein the composition is a spray dried.
 16. The composition and corresponding test analysis of claim 1, further comprising depolymerized chondroitin sulfate.
 17. The composition and corresponding test analysis of claim 1, further comprising at least 20% depolymerized chondroitin sulfate.
 18. A nutritional supplement that provides at least 500 mg of the composition of claim 1 in an effective daily amount.
 19. A nutritional supplement that provides at least 1000 mg of the composition of claim 1 in an effective daily amount.
 20. A nutritional supplement comprising a hard capsule containing at least some of the composition of claim
 1. 21. A nutritional supplement comprising a tablet containing at least some of the composition of claim
 1. 22. A nutritional supplement comprising at least some of the composition of claim 1 in a dosage form selected from the group consisting of a dispersible power, a granule, an aqueous suspension, an oil suspension, an emulsion, a soft capsule, a syrup and an elixir.
 23. A method of inducing cartilage formation in an individual with a connective tissue disorder, comprising orally administering to the individual an effective daily cartilage-inducing amount of the composition of claim
 1. 